García Yadiris, Vera Myleidi, González Duván, Vélez-Peña Estefanía, Sellergren Börje, Jiménez Verónica A
Departamento de Ciencias Químicas, Facultad de Ciencias Exactas, Universidad Andres Bello, Autopista Concepción-Talcahuano, Talcahuano 7100, Chile.
Departamento de Polímeros, Facultad de Ciencias Químicas, Universidad de Concepción, Edmundo Larenas 129, Concepción 4030000, Chile.
ACS Omega. 2025 May 13;10(20):20649-20660. doi: 10.1021/acsomega.5c01545. eCollection 2025 May 27.
Antibiotic determination at low concentrations is a critical challenge to ensure water, food, and environmental safety. This work reports the application of molecularly imprinted nanoparticles (MINs) as synthetic antibodies in a microplate assay for tetracycline (TC) determination at subnanomolar levels. MINs with hydrodynamic radii ranging from 200 to 300 nm were synthesized via solid-phase imprinting on magnetic nanoparticles using lymecycline (LMC) as an auxiliary template. MINs immobilized onto polystyrene microplates exhibited a selective binding of the conjugate between LMC and horseradish peroxidase (HRP), yielding analytical responses up to fourfold higher than those observed in blank wells. MINs saturated with LMC-HPR showed the quantitative displacement of the conjugate upon incubation with LMC or TC, yielding concentration-proportional responses from 0.001 to 100 nmol L. For LMC, we obtained limits of detection (LOD) between 0.018 and 0.020 nmol L and limits of quantification (LOQ) from 0.054 to 0.061 nmol L. For TC, the LOD values ranged between 0.001 and 0.006 nmol L, with LOQ values of 0.004-0.019 nmol L. Within the studied concentration range, the analytical responses of MINs were two to five times higher than those obtained from nonimprinted materials or blank controls. These findings support the performance of MINs as antibody mimetics with high recognition capacity, relatively low production costs, and robust analytical applications.
低浓度抗生素的测定是确保水、食品和环境安全的一项关键挑战。本研究报道了分子印迹纳米颗粒（MINs）作为合成抗体在微孔板分析中用于亚纳摩尔水平四环素（TC）测定的应用。通过使用赖甲环素（LMC）作为辅助模板在磁性纳米颗粒上进行固相印迹，合成了流体动力学半径在200至300nm之间的MINs。固定在聚苯乙烯微孔板上的MINs表现出对LMC与辣根过氧化物酶（HRP）之间共轭物的选择性结合，产生的分析响应比空白孔中观察到的高四倍。用LMC-HPR饱和的MINs在与LMC或TC孵育后显示共轭物的定量置换，产生0.001至100 nmol/L的浓度比例响应。对于LMC，我们获得的检测限（LOD）在0.018至0.020 nmol/L之间，定量限（LOQ）为0.054至0.061 nmol/L。对于TC，LOD值在0.001至0.006 nmol/L之间，LOQ值为0.004 - 0.019 nmol/L。在所研究的浓度范围内，MINs的分析响应比非印迹材料或空白对照获得的响应高两到五倍。这些发现支持了MINs作为具有高识别能力、相对较低生产成本和强大分析应用的抗体模拟物的性能。
